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In vitro development and cell allocation of porcine blastocysts derived by aggregation of in vitro fertilized embryos
Author(s) -
Lee SangGoo,
Park ChiHun,
Choi DonHo,
Kim HyeSun,
Ka HakHyun,
Lee ChangKyu
Publication year - 2007
Publication title -
molecular reproduction and development
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.745
H-Index - 105
eISSN - 1098-2795
pISSN - 1040-452X
DOI - 10.1002/mrd.20728
Subject(s) - biology , in vitro , embryo , microbiology and biotechnology , andrology , genetics , medicine
In pigs, the morphology and cell number of in vitro‐produced blastocysts are inferior to those of their in vivo counterparts. The objective of this study was to increase developmental competence and to gain an understanding of cell allocation in blastocysts derived from the aggregation of four‐cell stage porcine embryos produced in vitro. After removal of the zona pellucida, two (2×) and three (3×) four‐cell stage embryos were aggregated by co‐culturing them in aggregation plates. Five days after aggregation, the developmental ability and the number of cells in the aggregated embryos were determined. The percentage of blastocysts was higher ( P < 0.05) in both the 2× and 3× aggregated embryos (66.6% and 72.0%, respectively) compared to that of the 1× embryos and the intact controls (43.1% and 36.4%, respectively). The total cell number of blastocysts also increased in aggregated embryos compared to that of intact controls (2.6‐fold for 2× and 3.4‐fold for 3×) ( P < 0.05). The cells of two differentially stained embryos were started to mix at 72 hr after aggregation. In vitro‐fertilized porcine aggregates (2×) were developed to blastocyst with a random distribution of cells from each embryo. The mRNA levels for the oct‐4, bcl‐xL and connexin 43 genes were higher ( P < 0.05) and bak gene were lower ( P < 0.05) in both the 2× and 3× aggregated embryos than the intact controls. Therefore, the aggregation of the four‐cell stage embryos could be used to improve the quality of porcine preimplantation stage embryos produced in vitro. Mol. Reprod. Dev. 74: 1436–1445, 2007. © 2007 Wiley‐Liss, Inc.